Fiqriyana, MaulidyaAulia (2013) Studi Pengembangan Rapid Test Menggunakan Antibodi Poliklonal Hasil Induksi Autoantibodi TPO (Thyroidperoksidase) dari Serum Pasien Autoimun Tiroiditis dengan Metode Immunodot. Magister thesis, Universitas Brawijaya.
Abstract
Penyakit Autoimun Tiroid atau Autoimmune Thyroid Disease (AITD) adalah suatu penyakit kelainan autoimun yang ditandai dengan adanya inflamasi atau peradangan pada kelenjar tiroid. Inflamasi pada kelenjar tiroid menyebabkan aktivitas kelenjar tiroid dalam mensintesis hormon tiroid mengalami gangguan. Dalam sintesis hormon tiroid, enzim yang berperan adalah tiroid peroksidase (TPO). Pada penderita AITD, TPO tidak dikenali sebagai molekul self , tetapi dikenali sebagai non - self . Molekul non - self menyebabkan sistem imun menghasilkan autoantibodi yang akan melemahkan peran TPO sebagai molekul non - self tersebut. Konsentrasi serum yang mengandung autoantibodi TPO berkorelasi positif dengan aktivitas autoimun tiroiditis kronis. Oleh karena itu, autoantibodi TPO merupakan marker yang spesifik untuk deteksi dini AITD. Penelitian ini bertujuan untuk mengembangkan rapid test pada pasien AITD melalui tahapan isolasi dan karakterisasi autoantibodi TPO dan antibodi poliklonal terhadap autoantibodi TPO (anti-anti TPO). Serum pasien AITD diisolasi protein autoantibodi TPO dan dikonfirmasi pita proteinnya dengan metode WesternBlot. Dari hasil Westernblot diperoleh berat molekul TPO yaitu 51,81 kDa. Protein tersebut dikenali sebagai autoantibodi TPO, yang kemudian diimunisasikan ke kelinci ( Oryctolagus cuniculus ). Autoantibodi TPO dikenali sebagai antigen, sehingga tubuh kelinci menghasilkan respon imun berupa antibodi poliklonal. Serum antibodi poliklonal kelinci dikenali sebagai antibodi anti-TPO (anti-anti TPO) dan karakter antibodi poliklonal terhadap autoantibodi TPO (anti-anti TPO) dianalisis dengan ELISA, Dot Blotting dan Western blotting . Hasil penelitian menunjukkan bahwa dalam serum pasien AITD ditemukan marker autoantibodi TPO dengan berat molekul 51,8 kDa, marker dengan berat molekul 51,8 kDa digunakan untuk menginduksi poliklonal anti-anti TPO pada kelinci ( Oryctolagus cuniculus ). Kemudian, titer anti-anti TPO diukur dengan ELISA, sedangkan spesifitas dan sensitifitas ditentukan dengan dot blot dan Westernblot. Antibodi poliklonal hasil induksi autoantibodi TPO memiliki titer tertinggi pada bleeding ke-3 dengan absorbansi sebesar 0,3225 dan berat molekul 51,81 kDa yang mampu mengenali secara spesifik autoantibodi TPO.
English Abstract
Autoimmune thyroiditis disease (AITD) is an autoimmune disease that characterized by inflammatory of thyroid gland. inflammation can interfenfere shyntesis of thyroid. enzyme that plays a role in syn sis thyroid hormon is thyroid peroxidase (TPO), which is recognized as a non self molecul in AITD patients. Non self molecules cause immune responses in serum of AITD patients and will stimulate autoantibody production which weaken role of non self TPO. Concentration of autoantibody TPO in sera has a positive correlation with chronic AITD activities. refore, autoantibody TPO serum level is a spesific marker in early detection of AITD. aims of this research were to develop a rapid test in AITD patients by isolating and characterizing autoantibody TPO and polyclonal antibodies against autoantibody TPO (anti-anti TPO). autoantibody TPO was isolated from sera of AITD patients and band of proteins was confirmed by Westernblot. A molecular weight of autoantibody TPO confirmed by Westernblot method was 51.81 kDa, which was induced on rabbit ( Oryctolagus cuniculus ) and recognized as an antigen. In rabbit`s serum, polyclonal antibody was produced and identified as antibody anti TPO (anti-anti TPO), which was characterized by ELISA, dot blotting, and Western blotting. result showed that TPO antibody was found in serum as a marker, which molecular weight is 51.81 kDa. marker was used to induce polyclonal antibody anti-anti-TPO on rabbit ( Oryctolagus cuniculus ). titers of anti-anti-TPO were measured by ELISA for sensitivity test, while specitivity test were also measured by dot blot and Western blotting. Polyclonal Antibody induced by autoantibody TPO had highest titers on 3 rd serum bleeding within absorbance of 0.3225. result of dot blot and Westernblot showed that polyclonal antibody specifically recognized TPO autoantibody.
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/616.44/FIQ/s/041301370 |
| Subjects: | 600 Technology (Applied sciences) > 616 Diseases > 616.4 Diseases of endocrine, hematopoietic, lymphatic, glandular system; diseases of male breast |
| Divisions: | S2/S3 > Magister Matematika, Fakultas MIPA |
| Depositing User: | Endro Setyobudi |
| Date Deposited: | 05 Apr 2013 11:03 |
| Last Modified: | 05 Apr 2013 11:03 |
| URI: | http://repository.ub.ac.id/id/eprint/158354 |
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